Pharmaceutical co-crystal composition and use thereof

ABSTRACT

The current invention relates to series of co-crystals of platinum analogues and their pharmaceutical use. The co-crystals of the subject invention may be used in the treatment or prevention of cancers and virus infections.

FIELD OF THE INVENTION

The current invention relates to a series of co-crystals of platinumanalogues with diacids and the pharmaceutical use of these co-crystals.The co-crystals of the current invention may be used in the treatment orprevention of various diseases such as cancers and virus infections.

BACKGROUND OF THE INVENTION

The interest in platinum-based antitumor drugs has its origin in the1960's, with the serendipitous discovery by Rosenberg of the inhibitionof cell division by platinum (Pt) complexes. Since the approval ofcisplatin for the treatment of testicular and ovarian cancer in 1978,cisplatin has become one of the three most widely utilized antitumordrugs in the world. Platinum-based anticancer drugs have revolutionizedcancer chemotherapy, and continue to be in widespread clinical use,especially for management of tumors of the ovary, testes, and the headand neck. Thousands of Pt compounds have been synthesized and evaluatedas potential antitumor agents and over 28 have entered human clinicaltrials. However, several types of dose limiting toxicities associatedwith platinum drug use, partial anti-tumor response in most patients,development of drug resistance, tumor relapse, and other challenges haveseverely limited the patient quality of life. Therefore, it is desirableto develop new strategies for improving platinum therapy.

The search continues for an improved Pt antitumor agent. In the yearsfollowing the introduction of cisplatin, the design of new Pt antitumordrugs focused mainly on direct cisplatin analogues, which adhered to theset of structure-activity relationships summarized by Cleare andHoeschele in 1973. A number of researchers have taken a completelydifferent approach to Pt drug design and have produced compounds thatare inconsistent with the traditional structure-activity relationshipsbut still show antitumor activities.

Carboplatin, one of the second generation platin analogues, is lesstoxic than cisplatin and can be administered at a significantly higherdose than cisplatin (up to 2000 mg/dose); it has received worldwideapproval and has achieved routine clinical use. Unfortunately, thecontinued use of carboplatin is restricted by severe dose limiting sideeffects and intrinsic or acquired drug resistance.

In contrast to the 1970s and 1980s, the design of third-generation Ptdrugs in the recent years has clearly shifted away from the earlyempirical structure-activity relationships and the synthesis of merecisplatin analogues. Instead, efforts have been directed at the designof compounds capable of circumventing specific mechanisms of resistanceand at the design of unconventional Pt compounds with radicallydifferent modes of action. As the third-generation of compounds undergoclinical trials, it is hoped that they will demonstrate significantclinical advantages over the current drugs, particularly in the area ofPt drug resistance.

Meanwhile co-crystallization has attracted great amount of academic,industrial and therapeutic interests by co-crystallization of two ormore pure compounds with crystal engineering to create a new functionalmaterial. Specifically, pharmaceutical co-crystals are defined as“co-crystals in which the target molecule or ion is an activepharmaceutical ingredient, API, and it bonds to the co-crystal former(s)through hydrogen bonds.” Almarsson M. and Zaworotko J., Chem. Commun.,2004: 1889. Pharmaceutical co-crystals are nonionic supramolecularcomplexes and can be used to improve physiochemical properties such assolubility, stability and bioavailability in pharmaceutical developmentwithout changing the chemical composition of the active pharmaceuticalingredient (API).

Therefore, it is desirable to improve the physiochemical and therapeuticproperties of cisplatin, carboplatin and other platin withco-crystallization technology. In some cases, there is no need to changethe basic structure of the platin as the API, while properties such assolubility, stability, permeability and bioavailability can be improved.For example, it would be possible to significantly enhance thebioavailabiltiy of a platin API with co-crystallization, so that theco-crystal can be therapeutically effective in certain environment ofuse and maintain the level for a prolonged period of time.

The present invention provides a series of co-crystals including aplatinum analogue and a diacid as coformers. The co-crystals of thisinvention may satisfy one or more of the targeted objectives, such asbut not limited to increased solubility, stability and bioavailabilityand more versatility in pharmaceutical use.

SUMMARY OF THE INVENTION

The present invention provides a series of co-crystals comprising aplatinum analogue and a diacid. In some embodiments, co-crystals have astructure of Formula (I):

In some embodiments, the platinum analogue

is selected from the structures of formulas from Pt-00 to Pt-33 andSPI-77 listed in Tables 1-5. In some embodiments, the diacid

is selected from the structures of formulas CF-01, CF-02, CF-03, CF-04,CF-05, CF-06, CF-07, CF-08, CF-10A, CF-10B, CF-10C and CF-10D listed inTable 6.

In one aspect, the co-crystal of the present invention is formed wherethe platinum analogue, the active pharmaceutical ingredient (API), andthe diacid, the co-crystal former, are bonded together through hydrogenbonds. In some embodiments, other non-covalent interactions may also bein the co-crystal. In one embodiment, other non-covalent and covalentinteractions may also be present in the co-crystal.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising the compounds of the co-crystals of Formula I,wherein the co-crystal comprises a platinum analogue and a diacid. Insome embodiments of the pharmaceutical composition, the platinumanalogue is the API.

One aspect of the invention relates to platinum analogue-basedco-crystals which provide a sufficient level of bioavailability to betherapeutically effective in pharmaceutical use and maintains the levelfor a therapeutically effective period of time.

Another aspect of the invention is to provide uses of the compounds ofthe co-crystals (e.g. the co-crystals of Formal I) in certainindications; in some embodiments the uses of the compounds of theco-crystals extend beyond the uses of carboplatin by itself. In someembodiments, the present invention relates to treating or preventing adisease in a subject in need thereof comprising administering to thesubject the pharmaceutical composition comprising the compound of theco-crystal of Formula I, wherein the compound is in a therapeuticallyeffective amount. In some embodiments, the disease is a cancer; in otherembodiments, the disease is a virus infection.

In one aspect, the present invention involves the use of apharmaceutical composition comprising the compounds of the co-crystal ofthe current invention to induce cell death in cancer cells by contactingthe cancer cells with an effective amount of the compound.

In some embodiments of the treatment of cancers, the therapeuticallyeffective amount of the compound is about 0.01 to about 10 mg/kg bodyweight, and in some particular embodiments about 0.01 to about 5 mg/kgbody weight.

In some embodiments of treatment of virus diseases, the therapeuticallyeffective amount of the compound is about 0.01 to about 10 mg/kg bodyweight, and in some particular embodiments about 0.01 to 5 mg/kg bodyweight.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the IC₅₀ values of MD-39551 and the control chemicalsdocetaxel and cisplatin in PC-3 prostate cancer cell line.

FIG. 2 shows the IC₅₀ values of MD-39551 and the control chemicalsdocetaxel and cisplatin in LNCaP prostate cancer cell line.

FIG. 3 shows the IC₅₀ values of MD-39551 and the control chemicalsdocetaxel and cisplatin in fetal hepatocytes HL-7002.

FIG. 4 shows the IC₅₀ values of MD-39551 and the control chemicalsdocetaxel and cisplatin in human embryonic kidney cell line HEK293.

FIG. 5 shows the IC₅₀ values of MD-39703 and MD-39433 and the controlchemicals oxaliplatin and 5-FU in colorectal cancer cell line HCT-116.

FIG. 6 shows the IC₅₀ values of MD-39703 and MD-39433 and the controlchemicals oxaliplatin and 5-FU in colorectal cancer cell line HT-29.

FIG. 7 shows the IC₅₀ values of MD-39703 and MD-39433 and the controlchemicals oxaliplatin and 5-FU in fetal hepatocytes HL-7002.

FIG. 8 shows the IC₅₀ values of MD-39703 and MD-39433 and the controlchemicals oxaliplatin and 5-FU in human embryonic kidney cell lineHEK293.

FIG. 9 shows an X-ray powder diffraction (XRPD) pattern of theco-crystal MD-36042.

FIG. 10 shows a scanning electron microscope (SEM) image of theco-crystal MD-36042.

FIG. 11 shows an XRPD pattern of the co-crystal MD-39551.

FIG. 12 shows a differential scanning calorimetry (DSC) result of theco-crystal MD-39551.

FIG. 13 shows a SEM image of the co-crystal MD-39551.

FIG. 14 shows an XRPD pattern of the co-crystal MD-39442.

FIG. 15 shows an XRPD pattern of the co-crystal MD-39433.

FIG. 16 shows a SEM image of the co-crystal MD-39433.

FIG. 17 shows an XRPD pattern of the co-crystal MD-39703.

FIG. 18 shows a DSC result of the co-crystal MD-39703.

FIG. 19 shows a SEM image of the co-crystal MD-39703.

FIG. 20 shows an XRPD pattern of the co-crystal HP-309.

FIG. 21 shows an XRPD pattern of the co-crystal MD3176.

DETAILED DESCRIPTION OF THE INVENTION

The following description of certain embodiment(s) is merely exemplaryin nature and is in no way intended to limit the invention, itsapplication, or uses. As used throughout, ranges are used as shorthandfor describing each and every value that is within the range. Any valuewithin the range can be selected as the terminus of the range. Inaddition, all references cited herein are hereby incorporated byreferenced in their entireties. In the event of a conflict in adefinition in the present disclosure and that of a cited reference, thepresent disclosure controls. Unless defined otherwise, all technical andscientific terms used herein have the same meaning as is commonlyunderstood by one of skill in the art to which this invention belongs.All patents and publications referred to herein are incorporated byreference in their entireties.

The term “effective amount” or “therapeutically effective amount” refersto that amount of a compound or combination of compounds as describedherein that is sufficient to effect the intended application including,but not limited to, prophylaxis or treatment of diseases. Atherapeutically effective amount may vary depending upon the intendedapplication (in vitro or in vivo), or the subject and disease conditionbeing treated (e.g., the weight, age and gender of the subject), theseverity of the disease condition, the manner of administration, etc.which can readily be determined by one of ordinary skill in the art. Theterm also applies to a dose that will induce a particular response intarget cells and/or tissues (e.g., the reduction of cell proliferationand/or morphological alteration of the tissue). The specific dose willvary depending on the particular compounds chosen, the dosing regimen tobe followed, whether the compound is administered in combination withother compounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which the compound iscarried.

A “therapeutic effect” as that term is used herein, encompasses atherapeutic benefit and/or a prophylactic benefit. A “prophylacticeffect” (e.g. terms such as “prophylaxis,” “prevent” and “reducing thelikelihood for developing”) includes delaying or eliminating theappearance of a disease or condition, delaying or eliminating the onsetof symptoms of a disease or condition, slowing, halting, or reversingthe progression of a disease or condition, or any combination thereof byadministering a drug before the onset of the disease or condition. A“treatment effect” (e.g. with terms such as “treatment” and “treat”)includes reducing or eliminating the appearance of a disease orcondition, reducing or eliminating the symptoms of a disease orcondition, slowing, halting, or reversing the progression of a diseaseor condition, or any combination thereof by administering a drug afterthe onset of the disease or condition.

A “subject” as the term is used herein, refers to a human or non-humananimal. In some embodiments, the subject is a mammal. In someembodiments, the subject is human.

When ranges are used herein to describe, for example, physical orchemical properties such as molecular weight or chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. Use of the term “about” whenreferring to a number or a numerical range means that the number ornumerical range referred to is an approximation within experimentalvariability (or within statistical experimental error), and thus thenumber or numerical range may vary. In some embodiments, the variationis from 0% to 15%; in some particular embodiments from 0% to 10%; and inother embodiments from 0% to 5% of the stated number or numerical range.The term “comprising” (and related terms such as “comprise” or“comprises” or “having” or “including”) includes those embodiments suchas, for example, an embodiment of any composition of matter, method orprocess that “consist of” or “consist essentially of” the describedfeatures.

Compounds used in the present invention also include crystalline andamorphous forms of those compounds, including, for example, polymorphs,pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (includinganhydrates), conformational polymorphs, and amorphous forms of thecompounds, as well as mixtures thereof. “Compound of the co-crystal”refers to crystalline and amorphous forms made from the co-crystal,wherein “made from” means left unaltered or processed with known methodssuch as but not limited to dissolving, condensing, crystallinedisruption, drying, grinding, compaction, and polymer film coating.“Crystalline form” and “polymorph” are intended to include allcrystalline and amorphous forms of the compound, including, for example,polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs(including anhydrates), conformational polymorphs, and amorphous forms,as well as mixtures thereof, unless a particular crystalline oramorphous form is referred to.

The present invention relates to a series of co-crystals of platinumanalogues, and methods of making and using the same. The co-crystalcomprises a platinum analogue and a diacid as coformers. In someembodiments, the co-crystal formula is presented as Formula (I):

In some embodiments, the platinum analogue

represents the platinum-based anticancer drugs which are approved oralready in marketing shown in Table 1.

TABLE 1

Pt-00

Pt-01

Pt-02

Pt-03

Pt-04

Pt-05

Pt-06

In some embodiments, the platinum analogue

represents lipoplatin and other platinum-based anticancer drugs shown inTable 2.

TABLE 2

Pt-07

Pt-08

In some embodiments, the platinum analogue

represents the platinum-based anticancer drugs in clinic phases shown inTable 3.

TABLE 3

Pt-09

Pt-10

Pt-11

Pt-12

Pt-13

Pt-14

Pt-15

Pt-16

In some embodiments, the platinum analogue

represents the platinum-based anticancer drugs in study as shown inTable 4.

TABLE 4

Pt-17

Pt-18

Pt-19

Pt-20

In some embodiments, the platinum analogue

represents the platinum-based structures as shown in Table 5.

TABLE 5

Pt-21

Pt-22

Pt-23

Pt-24

Pt-25

Pt-26

Pt-27

Pt-28

Pt-29

Pt-30

Pt-31

In some embodiments, the platinum analogue

represents the platinum-based anticancer drugs JM11 and iproplatin asdisclosed in Wheate, S. et al. Dalton Trans., 2010, Vol. 39, 8113-27.The structure of JM11 and iproplatin are shown below:

In some embodiments of Formula (I), n is an integer selected from 0 to5; in some embodiments of Formula (I), n is an integer selected from 2to 5. In some embodiments, when n≥2, the carbon atoms on the diacid maybe connected by single or double bonds.

In some embodiments, the platinum analogue and the diacid are bonded ata 1:1 ratio.

In some embodiments, R₅ and R₆ are the same as or different from eachother, and independently represent a hydrogen, a halogen, an aminogroup, a C1-C6 alkyl group, a cyanide group, a hydroxyl group, an acylgroup, a phosphoryl group, a phosphoroamido group, a hydroxylcarboxylgroup, a phenyl group, or an aliphatic group, or R5 and R6 are connectedto form a substituted or unsubstituted C3-C6 cycloalkyl group or phenylgroup.

In some embodiments, the diacid

may be selected from the group consisting of oxalic acid,1,3-propanedioic acid, 1,4-butanedioic acid, 1,5pentanedioic acid,cis-butenedioic acid, 2-hydroxy-1,4-butanedioic acid (malic acid),2,3-dihydroxy-1,4-butanedioic acid (tartaric acid),2-phenyl-1,3-propanedioic acid, 1,2-dicarboxycyclohexane,3-hydroxy-3-carboxy-1,5-pentanedioic acid (citric acid), phthalic acid,and 1,3,4-benzene-ticarboxylic acid.

In some embodiments, the diacid

may be selected from formulas CF-01, CF-02, CF-03, CF-04, CF-05, CF-06,CF-07, CF-08, CF-10A, CF-10B, CF-10C and CF-10D in Table 6.

TABLE 6 CF-09

CF-10

CF-01

CF-02

CF-03

CF-04

CF-05

CF-06

CF-07

CF-08

CF-10A

CF-10B

CF-10C

CF-10D

When the platinum analogue is carboplatin (Pt-01), co-crystals disclosedin U.S. Pat. App. Pub. No. 20050160931 and U.S. Pat. No. 6,699,901 arenot included in the co-crystals of the present invention. In particular,when the platinum analogue is carboplatin (Pt-01), the diacid does nothave the structure of CF-01, CF-04, CF-10A, CF-10B, CF-10C, CF-10D asshown in Table 6.

In some embodiments, the co-crystal of the present invention comprises aplatinum analogue selected from the group consisting of formulas Pt-01,Pt-02, Pt-03 and Pt-05.

In some embodiments, the co-crystal of the present invention comprises adiacid selected from the group consisting of formulas CF-01, CF-02,CF-08.

In some embodiments, the co-crystal of the present invention comprises aplatinum analogue selected from the group consisting of formulas Pt-01,Pt-02, Pt-03 and Pt-05 and a diacid selected from the group consistingof formulas CF-01, CF-02, CF-08.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-01 and the diacid of formula CF-02bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 8.821°,8.961°, 11.998°, 13.160°, 17.681°, 18.001°, 19.101° and 20.837° (roundto 8.8°, 9.0°, 12.0°, 13.2°, 17.7°, 18.0°, 19.1° and 20.8°,respectively) (corresponding to d-spacing of 10.0166 Å, 9.8604 Å, 7.3703Å, 6.7219 Å, 5.0120 Å, 4.9237 Å, 4.6427 Å and 4.2595 Å, respectively)±0.2. In some embodiments, the co-crystal has an XRPD pattern comprisingpeaks at diffraction angles 2-Theta of 8.821°, 8.961°, 11.998°, 13.160°,17.681°, 18.001°, 19.101° and 20.837°±0.1. In some embodiments, theco-crystal has an XRPD pattern comprising peaks at diffraction angles2-Theta of 8.821°, 8.961°, 11.998°, 13.160°, 17.681°, 18.001°, 19.101°and 20.837°±0.05. In some embodiments, the co-crystal has an x-raydiffraction pattern comprising peaks as set forth in FIG. 9. In someembodiments, the co-crystal has an x-ray diffraction patternsubstantially similar to the pattern as set forth in FIG. 9.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-01 and the diacid of formula CF-08bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 6.338°,14.437°, 14.860°, 15.281°, 19.958°, 22.682° and 24.600° (round to 6.3°,14.4°, 14.9°, 15.3°, 20.0°, 22.7° and 24.6°, respectively)(corresponding to d-spacing of 13.9342 Å, 6.1301 Å, 5.9567 Å, 5.7936 Å,4.4451 Å, 3.9171 Å and 3.6158 Å respectively) ±0.2. In some embodiments,the co-crystal has an XRPD pattern comprising peaks at diffractionangles 2-Theta of 6.338°, 14.437°, 14.860°, 15.281°, 19.958°, 22.682°and 24.600°±0.1. In some embodiments, the co-crystal has an XRPD patterncomprising peaks at diffraction angles 2-Theta of 6.338°, 14.437°,14.860°, 15.281°, 19.958°, 22.682° and 24.600°±0.05. In someembodiments, the co-crystal has an x-ray diffraction pattern comprisingpeaks as set forth in FIG. 11. In some embodiments, the co-crystal hasan x-ray diffraction pattern substantially similar to the pattern as setforth in FIG. 11.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-02 and the diacid of formula CF-02bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.338°,9.401°, 10.057°, 12.535°, 13.619° and 23.361° (round to 7.3°, 9.4°,10.1°, 12.5°, 13.6° and 23.4°, respectively) (corresponding to d-spacingof 12.0363 Å, 9.3993 Å, 8.7877 Å, 7.0557 Å, 6.4967 Å and 3.8048 Årespectively) ±0.2. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.338°,9.401°, 10.057°, 12.535°, 13.619° and 23.361°±0.1. In some embodiments,the co-crystal has an XRPD pattern comprising peaks at diffractionangles 2-Theta of 7.338°, 9.401°, 10.057°, 12.535°, 13.619° and23.361°±0.05. In some embodiments, the co-crystal has an x-raydiffraction pattern comprising peaks as set forth in FIG. 14. In someembodiments, the co-crystal has an x-ray diffraction patternsubstantially similar to the pattern as set forth in FIG. 14.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-02 and the diacid of formula CF-01bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.079°,9.180°, and 10.060° (round to 7.1°, 9.2°, and 10.1°, respectively)(corresponding to d-spacing of 12.4769 Å, 9.6252 Å and 8.7856 Årespectively) ±0.2. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.079°,9.180°, and 10.060°±0.1. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.079°,9.180°, and 10.060°±0.05. In some embodiments, the co-crystal has anx-ray diffraction pattern comprising peaks as set forth in FIG. 15. Insome embodiments, the co-crystal has an x-ray diffraction patternsubstantially similar to the pattern as set forth in FIG. 15.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-02 and the diacid of formula CF-08bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.858°,11.881°, 14.463°, 15.757°, 16.999°, 17.376° and 17.841° (round to 7.9°,11.9°, 14.5°, 15.8°, 17.0°, 17.4° and 17.8°, respectively)(corresponding to d-spacing of 11.2418 Å, 7.4427 Å, 6.1193 Å, 5.6194 Å,5.2115 Å, 5.0993 Å and 4.9676 Å respectively) ±0.2. In some embodiments,the co-crystal has an XRPD pattern comprising peaks at diffractionangles 2-Theta of 7.858°, 11.881°, 14.463°, 15.757°, 16.999°, 17.376°and 17.841°±0.1. In some embodiments, the co-crystal has an XRPD patterncomprising peaks at diffraction angles 2-Theta of 7.858°, 11.881°,14.463°, 15.757°, 16.999°, 17.376° and 17.841°±0.05. In someembodiments, the co-crystal has an x-ray diffraction pattern comprisingpeaks as set forth in FIG. 17. In some embodiments, the co-crystal hasan x-ray diffraction pattern substantially similar to the pattern as setforth in FIG. 17.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-03 and the diacid of formula CF-01bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 7.181°,9.499°, 13.740° and 14.421° (round to 7.2°, 9.5°, 13.7° and 14.4°,respectively) (corresponding to d-spacing of 12.2995 Å, 9.3029 Å, 6.4398Å and 6.1371 Å respectively) ±0.2. In some embodiments, the co-crystalhas an XRPD pattern comprising peaks at diffraction angles 2-Theta of7.181°, 9.499°, 13.740° and 14.421°±0.1. In some embodiments, theco-crystal has an XRPD pattern comprising peaks at diffraction angles2-Theta of 7.181°, 9.499°, 13.740° and 14.421°±0.05. In someembodiments, the co-crystal has an x-ray diffraction pattern comprisingpeaks as set forth in FIG. 20. In some embodiments, the co-crystal hasan x-ray diffraction pattern substantially similar to the pattern as setforth in FIG. 20.

In some embodiments, the co-crystal of the present invention comprisesthe platinum analogue of formula Pt-05 and the diacid of formula CF-01bonded at 1:1 ratio. In some embodiments, the co-crystal has an XRPDpattern comprising peaks at diffraction angles 2-Theta of 6.697°,7.381°, 8.239°, 12.320° and 16.478° (round to 6.7°, 7.4°, 8.2°, 12.3°and 16.5°, respectively) (corresponding to d-spacing of 13.1874 Å,11.9662 Å, 10.7224 Å, 7.1785 Å and 5.3751 Å respectively) ±0.2. In someembodiments, the co-crystal has an XRPD pattern comprising peaks atdiffraction angles 2-Theta of 6.697°, 7.381°, 8.239°, 12.320° and16.478°±0.1. In some embodiments, the co-crystal has an XRPD patterncomprising peaks at diffraction angles 2-Theta of 6.697°, 7.381°,8.239°, 12.320° and 16.478°±0.05. In some embodiments, the co-crystalhas an x-ray diffraction pattern comprising peaks as set forth in FIG.21. In some embodiments, the co-crystal has an x-ray diffraction patternsubstantially similar to the pattern as set forth in FIG. 21.

In some embodiments, the co-crystal of the present invention comprises:(i) a diacid as a co-former; and (ii) a platinum analogue as a co-formerand the active pharmaceutical ingredient (API). In some embodiments, thediacid and the platinum analogue are bonded in 1:1 ratio.

As described here, the solid state of the co-crystal of the currentinvention is any crystalline polymorphic forms or a mixture thereof. Theco-crystal may also be made into an amorphous form, which may becombined with any crystalline forms. In other embodiments, the solidstate of the co-crystal is an amorphous form. Different forms of theco-crystal of the current invention may be obtained through differentcrystallization process and the co-crystals may be made into amorphousforms with known technology.

The compound of the co-crystals of the current invention (e.g.co-crystals of formula I) may demonstrate a sufficient level ofbioavailablity to be therapeutically effective in pharmaceutical use andmaintains that level in a subject for a prolonged period of time.

The co-crystals of the current invention may be produced by a processcomprising: (i) providing and mixing a platinum analogue, a diacid andan appropriate solvent, (ii) slurrying or stirring the mixture from stepi) for a sufficient period of time; and (iii) isolating the co-crystalformed thereby. In some embodiments, the reaction of the platinumanalogue and the diacid may be carried out 30° C. In some embodiments,the mixture after the reaction may be cooled to 0-5° C. and stirred.

The specific conditions of the process may be adjusted to ensureoptimized purity, quantity, and/or physiochemical properties. In someembodiments, the proper ratio is in the molar range of 1:0.1-1:20,1:0.2-1:20, 1:0.3-1:20, 1:0.4-1:20, 1:0.5-1:20, 1:0.6-1:20, 1:0.7-1:20;1:0.8-1:20, 1:0.9-1:20, 1:1-1:1.20, 1:2-1:20, 1:3-1:20, 1:4-1:20,1:5-1:20, 1:6-1:18, 1:7-1:15, 1:8-1:13, 1:9-1:12, or 1:10-1:11. In someembodiments, the proper ratio is about 1:1 (molar). In some embodiments,the period of time for slurrying or stirring the mixtures may be in therange of 0.1-24 hours, 0.2-12 hours, 0.25-6 hours, 0.3-2 hours, 0.4-1hour, or 0.5-1 hour. In some embodiments, the period of time forslurrying or stirring the mixtures may be about 0.5 hour. In someembodiments, the co-crystal compound may be obtained by drying,filtering, centrifugation, pipetting, or a combination thereof. In someembodiments, the co-crystal compound may be obtained by centrifugation.

In some embodiments, the reaction of the platinum analogue and thediacid may be carried out 30° C. In some embodiments, the mixture afterthe reaction may be cooled to 0-5° C. and stirred.

The current invention relates to the pharmaceutical use of compounds ofthe co-crystals of the present invention, and methods of treating orpreventing a disease in a subject in need thereof. In some embodiments,the method comprises administering to the subject a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof one or more of the co-crystals of the present invention.

In some embodiments, the compound of the co-crystal of the currentinvention demonstrates advantageous therapeutic properties. For example,in some embodiments, the compound of the co-crystals of the presentinvention may be more effective in killing cancerous or virus-infectedcells compared to carboplatin or other known drugs. In otherembodiments, the compound of the co-crystals of the present inventionmay be less effective in killing cancerous or virus-infected cellscompare to carboplatin or other known drugs or have substantiallysimilar effects, but are less toxic to healthy and normal cells,resulting in a net health benefit. For instance, comparing to knowplatin analogues in the treatment of cancer cells or virus-infectedcells, a compound of the MD39551 (as shown in Table 7) co-crystal may beless toxic and more stable than cisplatin and carboplatin. In addition,a compound of the MD39433 or MD39703 (as shown in Table 7) co-crystalsmay be less toxic and more stable than oxaliplatin and carboplatin. Insome embodiments, the compounds of MD39551, MD39433 or MD39703 mayprovide reduced side effects. In some embodiments, the compounds ofMD39551, MD39433 or MD39703 may demonstrate more versatility inpharmaceutical uses, e.g. when compared to carboplatin.

In some embodiments, the compound of the carboplatin-based co-crystal ofthe current invention demonstrates advantageous physiochemicalproperties. For example, in some embodiments, compounds of MD39551,MD39433 or MD39703 may have increased solubility, stability, andbioavailability. For example, in comparison with carboplatin, thecompounds of MD39551, MD39433 or MD39703 may be more stable and could bestable in solid form of various doses. Meanwhile, water solubility ofcompounds of MD39551, MD39433 or MD39703 may be higher than carboplatin,providing significantly more possibility of formulations andadministration.

In some embodiments, the pharmaceutical composition may consist of thecompounds of the co-crystals of the present invention. In someembodiments, the pharmaceutical composition may comprise the compoundsof the co-crystals of the present invention and at least one additionaltherapeutic agent or adjuvant therapy agent. The additional therapeuticagent or adjuvant therapy agent may be selected from but is not limitedto: folic acid, coenzyme Q10, curcumin, glutathione (GSH), aloe vera,oryzanol, 5-fluorouracil, bortezomib, or a combination thereof.Depending on the particular disease to be treated, the additionaltherapeutic agent or adjuvant therapy agent may include drugs alreadyknown. In some embodiments, the additional therapeutic agent or adjuvanttherapy agent may include drugs that have already been clinicallyaccepted to treat or prevent the disease.

In some embodiments, the pharmaceutical composition may comprise thecompounds of the co-crystals of the present invention and apharmaceutically acceptable carrier or excipient. “Pharmaceuticallyacceptable carrier” or “pharmaceutically acceptable excipient” isintended to include any and all solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, and inert ingredients. The use of such pharmaceuticallyacceptable carriers or pharmaceutically acceptable excipients for activepharmaceutical ingredients is well known in the art. Except insofar asany conventional pharmaceutically acceptable carrier or pharmaceuticallyacceptable excipient is incompatible with the active pharmaceuticalingredient, its use in the therapeutic compositions of the invention iscontemplated. Additional active pharmaceutical ingredients, such asother drugs, can also be incorporated into the described compositionsand methods.

In yet another aspect, the amount of the compound of the co-crystals ofthe present invention in the pharmaceutical composition administered toa subject may be about 0.005 to 20 mg/kg body weight, about 0.005 to 10mg/kg body weight, about 0.005 to 5 mg/kg body weight, about 0.005 to2.5 mg/kg body weight, 0.01 to 20 mg/kg body weight, about 0.01 to 10mg/kg body weight, about 0.01 to 5 mg/kg body weight, about 0.01 to 2.5mg/kg body weight, 0.1 to 20 mg/kg body weight, about 0.1 to 10 mg/kgbody weight, about 0.1 to 5 mg/kg body weight, or about 0.1 to 2.5 mg/kgbody weight. The specific amount of the compound depends on theparticular disease to be treated and the subject's specific conditions.

In yet another aspect, the administration of the pharmaceuticalcomposition comprising the compounds of the co-crystals of the presentinvention may last at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84, 91 or 98 days. In someembodiments, the administering of the pharmaceutical composition maylast at least one week. In some embodiments, the administering of thepharmaceutical composition may last at least two weeks. The specificperiod of administration depends on the particular disease to be treatedand the subject's specific conditions.

The present invention in various aspects and embodiments involves usesof the co-crystals of the present invention for the prevention ortreatment of various diseases and methods of treating or preventing thediseases by administering a pharmaceutical composition comprising thecompounds of the co-crystals of the present invention. The diseases tobe treated or prevented include but are not limited to cancers and viralinfections. For example, the co-crystals may be MD39551, MD39433 orMD39703.

In some embodiments, the disease is a cancer. In some embodiments, thecancer is selected from: bladder cancer, non-small cell lung cancer,cervical cancer, anal cancer, pancreatic cancer, squamous cell carcinomaincluding head and neck cancer, renal cell carcinoma, basal-cell skincancer (BCC), squamous-cell skin cancer (SCC), melanoma, ovarian cancer,small cell lung cancer, endometrial cancer, glioblastoma, astroycytoma,oligodendroglioma, ependymoma, neurofibrosarcoma, meningioma,gastrointestinal stromal tumor, breast cancer, lung cancer, colorectalcancer, thyroid cancer, bone sarcoma, stomach cancer, oral cavitycancer, oropharyngeal cancer, gastric cancer, renal adenocarcinoma,liver cancer, prostate cancer, esophageal cancer, testicular cancer,gynecological cancer, colorectal cancer, brain cancer, leukemia,leucocythemia, chronic lymphocytic leukemia (CLL), small lymphocyticleukemia (SLL), non-Hodgkin's lymphoma (NHL), diffuse large B celllymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL),Hodgkin's lymphoma, B cell acute lymphoblastic leukemia (B-ALL),Burkitt's lymphoma, Waldenström's macroglobulinemia (WM), Burkitt'slymphoma, multiple myeloma, and myelofibrosis.

In some embodiments, the pharmaceutical composition comprising thecompound of the co-crystals of the present invention may be used toprevent or treat prostate cancer, colorectal cancer, or renaladenocarcinoma. In some embodiments, the therapeutically effectiveamount of the co-crystals of the present invention to prevent or treatcancer may about 0.01 to about 10 mg/kg body weight. In anotherembodiment, the therapeutically effective amount of the compound of theco-crystals of the present invention to prevent or treat cancer is about0.01 to about 5 mg/kg body weight.

In some embodiments, the disease is a viral infection. In someembodiments, the virus is a DNA virus or an RNA virus. For example, insome embodiments the virus may be a DNA virus such as but not limited toadenovirus, herpes simplex virus, human pepillomavrus, VITAMIN K virus,smallpox virus, hepatitis B virus (HBV), and parvovirus B19. In otherembodiments, the virus may be an RNA virus such as but not limited tohuman astrovirus, norwalk virus, hepatitis A virus (HAV), severe acuterespiratory syndrome virus, hepatitis C virus (HCV), yellow fever virus,dengue virus, West Nile virus, TBE virus, rubella virus, hepatitis Evirus (HEV), human immunodeficiency virus (HIV), influenza virus, Lassavirus (LASV), Crimean-Congo hemorrhagic fever virus, Hantaan virus,Ebola virus, Marburg virus, Measles virus, mumps virus, parainfluenzavirus, respiratory syncytial virus, rabies virus, and hepatitis D virus(HDV), rotavirus, orbivirus, coltivirus, Banna virus.

In some embodiments, the pharmaceutical composition may be used toprevent or treat viral infections caused by HBV, HCV, HIV or Hantaanvirus. In some embodiments, the therapeutically effective amount of thecompound of the co-crystals of the present invention to prevent or treatviral infection is about 0.01 to about 10 mg/kg body weight. In anotherembodiment, the therapeutically effective amount of the compound of theco-crystals of the present invention to prevent or treat cancer is about0.01 to about 5 mg/kg body weight.

In some embodiments, the present invention provides a method oftreating, preventing, reducing or alleviating the symptoms of, and/orslowing or halting the progress of prostate cancer, colorectal cancer,renal adenocarcinoma or leucocythemia in a subject in need thereof, themethod comprising administrating to the subject an effective amount of apharmaceutical composition comprising the compound of the co-crystals ofthe present invention. In some embodiments, the pharmaceuticalcomposition consists of the compound of the co-crystals of the presentinvention. In some embodiments, the pharmaceutical composition furthercomprises at least one additional therapeutic agent or adjuvant therapyagent. In a specific embodiment, the additional therapeutic agent oradjuvant therapy agent may be selected from: folic acid, coenzyme Q10,curcumin, glutathione (GSH), aloe vera, oryzanol, 5-fluorouracil, andbortezomib. In some embodiments, the pharmaceutical compositioncomprises the compound of the co-crystals of the present invention and apharmaceutically acceptable carrier or excipient.

In some embodiments, the present invention provides a method oftreating, preventing, reducing or alleviating the symptoms of, and/orslowing or halting the progress of prostate cancer, the methodcomprising administrating to the subject an effective amount of apharmaceutical composition comprising the compound of co-crystalMD39551.

In some embodiments, the present invention provides a method oftreating, preventing, reducing or alleviating the symptoms of, and/orslowing or halting the progress of colorectal cancer, the methodcomprising administrating to the subject an effective amount of apharmaceutical composition comprising the compound of co-crystal MD39433or MD39703.

In some embodiments, the present invention provides a method oftreating, preventing, reducing or alleviating the symptoms of, and/orslowing or halting the progress of viral infections caused by HBV, HCV,HIV or Hantaan virus in a subject in need thereof, the method comprisingadministrating to the subject an effective amount of a pharmaceuticalcomposition comprising the compound of the co-crystals of the presentinvention. In some embodiments, the pharmaceutical composition consistsof the compound of the co-crystals of the present invention. In someembodiments, the pharmaceutical composition further comprises at leastone additional therapeutic agent or adjuvant therapy agent. In aspecific embodiment, the additional therapeutic agent or adjuvanttherapy agent may be selected from: folic acid, coenzyme Q10, curcumin,glutathione (GSH), aloe vera, oryzanol, 5-fluorouracil, and bortezomib.In some embodiments, the pharmaceutical composition comprises thecompound of the co-crystals of the present invention and apharmaceutically acceptable carrier or excipient.

In some embodiments, the administration of the pharmaceuticalcomposition according to the present invention can be via any commonroute as long as the target issue is available via the route. Suitableroutes may include oral, buccal, by inhalation spray, sublingual,rectal, transdermal, vaginal, transmucosal, topical, nasal or intestinaladministration; parenteral delivery, including intramuscular,subcutaneous, intramedullary injections, as well as intrathecal, directintraventricular, orthotopic, intrademal, intraperitoneal, intravenous,intra-articular, intra-sternal, intra-synovial, intra-hepatic,intralesional, intracranial, intraperitoneal, intranasal, or intraocularinjections or other modes of delivery. The preferred delivery routedepends on the particular disease to be treated and the subject'sspecific conditions.

In some embodiments, for prevention or treatment of prostate cancer,colorectal cancer, renal adenocarcinoma or leucocythemia, thepharmaceutical composition comprising the compound of the co-crystals ofthe present invention is administered with infusion, injections or viathe oral route. In some embodiments, for prevention or treatment ofprostate cancer, colorectal cancer, renal adenocarcinoma orleucocythemia, the pharmaceutical composition comprising the compound ofthe co-crystals of the present invention is administered for at leastone, two or three weeks.

In some embodiments, for prevention or treatment of viral infectionscaused by HBV, HCV, HIV or Hantaan virus, the pharmaceutical compositioncomprising the compound of the co-crystals of the present invention isadministered with infusion, injections or via the oral route. In someembodiments, for prevention or treatment of viral infections caused byHBV, HCV, HIV or Hantaan virus, the pharmaceutical compositioncomprising the compound of the co-crystals of the present invention isadministered for at least one, two or three weeks.

EXAMPLES

The effects of the co-crystal of the present invention on certaindiseases are shown in the following example. In addition, the process ofmaking the co-crystals of the present invention and the physiochemicalproperties of these crystals are also described. These examples do notin any way limit the scope of the invention.

A number of co-crystals are produced by mixing a platinum analogue witha diacid. The resulting co-crystals meet partly or completely thetargeted objectives, such as increased solubility, stability andbioavailability and more versatile in pharmaceutical use compared tocarboplatin or other platin compounds.

In comparison with carboplatin, the co-crystal of the current inventionsis more stable and can be stable in solid forms. In comparison to thereported platin analogues for the treatment of cancer cells, some of theco-crystals of the current inventions are less toxic and much stablethan cisplatin and carboplatin.

The inventors have determined that the formation of crystallinepolymorphic forms was confirmed with various methods such as but notlimited to XRPD, HPLC, ¹H-NMR; DSC and SEM. Amorphous forms of theco-crystal and other forms may be existent using differentcrystallization process.

The Effects of MD-39551 on Prostate Cancer Cells

The co-crystal MD-39551, which comprises the platinum analogue offormula Pt-01 and the diacid of formula CF-08 bonded at 1:1 ratio, wastested in the treatment of prostate cancers and compared to docetaxeland cisplatin, widely accepted drugs for prostate cancer patients.

PC-3 cells are a cell line derived from advanced prostate cancer patientwith bone metastasis and are characteristic of prostate cancer such asprostate small cell carcinoma. PC-3 cells were treated with drugs(MD-39551, docetaxel, or cisplatin) at step-wise concentrations, and thecell viability was evaluated with the CellTiter 96 AQueous One SolutionCell Proliferation Assay from Promega Corp. (Madison, Wis., USA). Theindex of cell growth repression ratio was obtained by comparing theOD490 data of treatment group to the negative control. The drug responserate IC₅₀ was calculated with the SPSS 16.0 system. The results areshown in FIG. 1.

The IC₅₀ of MD-39551 was 47.186 μM, while IC₅₀ of docetaxel andcisplatin were 49.924 μM and 2.489 μM respectively (FIG. 1).

LNCaP cells are a cell line derived from advanced prostate cancerpatient with lymph node metastasis. LNCaP cells were treated with drugs(MD-39551, docetaxel, or cisplatin) at step-wise concentrations, and thecell viability was evaluated with the CellTiter 96 AQueous One SolutionCell Proliferation Assay from Promega Corp. (Madison, Wis., USA). Theindex of cell growth repression ratio was obtained by comparing theOD490 data of treatment group to the negative control. The drug responserate IC₅₀ was calculated with the SPSS 16.0 system. The results areshown in FIG. 2.

For LNCaP cells, the IC₅₀ of MD-39551 was 33.232 μM; the IC_(50s) ofdocetaxel and cisplatin were 4.034 μM and 2.245 μM respectively (FIG.2).

HL-7002 cells are an immortalized human fetal hepatic cell line. HL-7002cells were treated with drugs (MD-39551, docetaxel, or cisplatin) atstep-wise concentrations, and the cell viability was evaluated with theCellTiter 96 AQueous One Solution Cell Proliferation Assay from PromegaCorp. (Madison, Wis., USA). The index of cell growth repression ratiowas obtained by comparing the OD490 data of treatment group to thenegative control. The drug response rate IC₅₀ was calculated with theSPSS 16.0 system. The results are shown in FIG. 3.

For HL-7002 cells, while no MD-39551 toxicity was detected, docetaxeland cisplatin showed significant toxicity. The IC₅₀ of docetaxel andcisplatin were 0.095 μM and 2.008 μM respectively (FIG. 3).

HEK293 cells are an immortalized human fetal kidney cell line. HEK293cells were treated with drugs (MD-39551, docetaxel, or cisplatin) atstep-wise concentrations, and the cell viability was evaluated with theCellTiter 96 AQueous One Solution Cell Proliferation Assay from PromegaCorp. (Madison, Wis., USA). The index of cell growth repression ratiowas obtained by comparing the OD490 data of treatment group to thenegative control. The drug response rate IC₅₀ was calculated with theSPSS 16.0 system. The results are shown in FIG. 4.

For HEK293 cells, while no MD-39551 toxicity was detected, docetaxel andcisplatin showed significant toxicity. The IC₅₀ of docetaxel andcisplatin were 1.741 μM and 6.899 μM, respectively (FIG. 4.).

Methods and Strategies:

Cell culture: Prostate cancer cell lines LNCaP and PC-3 were purchasedfrom ATCC (Manassas, Va.). The fetal hepatocytes HL-7002 and humanembryonic kidney cells HEK393 were purchased from ATCC. The cells werecultured in RPMI+5% Fetal Bovine Serum (FBS).

Drug treatment and cell viability (MTS) assay: The cells (105/100mL/well) were cultured in a 96 well plate, and treated with drugs (e.g.MD-39551) at step-wise concentrations from 0.01 to 300 μM. The cellstreated with the solvents were used as the negative control, andcisplatin and docetaxel were used as the positive controls. The cellswere monitored daily, and the cell viability was evaluated with thePromega CellTiter 96 AQueous One Solution Cell Proliferation Assay(Promega, Madison, Wis., USA) according to the manufacture manuals. Thecell viability was monitored at OD490 reading in a bio-spectrometer(Perkin Elmer, Walthan, Mass., USA).

Data analysis: The OD490 reading data were collected hourly from 1 h to4 h after the addition of lysis buffer. The index of cell growthrepression ratio was obtained by comparing the OD490 data of treatmentto the negative control. The drug response rate IC₅₀ was calculated withthe SPSS 16.0.

Summary of Effects:

For PC-3, a cell line derived from advanced prostate cancer patient withbone metastasis, MD-39551 showed similar cellular toxicity to docetaxel,but weaker than for Cisplatin. For LNCaP, a cell line derived fromadvanced prostate cancer patient with lymph node metastasis, thecellular toxicity of MD-39551 was weaker than docetaxel and cisplatin.For HL-7002 and HEK293 cells, which respectively represent normal humanhepatic cells and kidney cells, while MC-39551 showed no toxicity, bothdocetaxel and cisplatin demonstrated high toxicity.

The Effects of MD-39703 and MD-39433 on Colorectal Cancer Cells

The co-crystals MD-39703 and MD-39433 was tested in the treatment ofcolorectal cancers in comparison to oxaliplatin and fluorouracil (5-FU),widely used drugs in treating colorectal cancer patients. MD-39703comprises the platinum analogue of formula Pt-02 and the diacid offormula CF-08 bonded at 1:1 ratio; MD-39433 comprises the platinumanalogue of formula Pt-02 and the diacid of formula CF-01 bonded at 1:1ratio.

HCT-116 cells are a colorectal cancer cell line. HCT-116 cells weretreated with drugs (MD-39703, MD-39433, oxaliplatin, or 5-FU) atstep-wise concentrations, and the cell viability was evaluated with theCellTiter 96 AQueous One Solution Cell Proliferation Assay from PromegaCorp. (Madison, Wis., USA). The index of cell growth repression ratiowas obtained by comparing the OD490 data of treatment group to thenegative control. The drug response rate IC₅₀ was calculated with theSPSS 16.0 system. The results are shown in FIG. 5.

For the reduction of HCT-116 cell number, the IC₅₀ of MD-39703 was26.019 μM and IC₅₀ of MD-39433 was 25.293; IC_(50s) of oxaliplatin and5-FU were be 8.151 μM and 4.214 μM respectively (FIG. 5).

HT29 cells are a colorectal cancer cell line. HT29 cells were treatedwith drugs (MD-39703, MD-39433, oxaliplatin, or 5-FU) at step-wiseconcentrations, and the cell viability was evaluated with the CellTiter96 AQueous One Solution Cell Proliferation Assay from Promega Corp.(Madison, Wis., USA). The index of cell growth repression ratio wasobtained by comparing the OD490 data of treatment group to the negativecontrol. The drug response rate IC₅₀ was calculated with the SPSS 16.0system. The results are shown in FIG. 6.

For HT29 cells, the IC₅₀ of MD-39703 was 24.865 μM and IC₅₀ of MD-39433was 24.941; IC₅₀, of oxaliplatin and 5-FU were determined to be 29.993μM and 7.556 respectively (FIG. 6).

HL-7002 hepatocyte cell line cells were treated with drugs (MD-39703,MD-39433, oxaliplatin, or 5-F U) at step-wise concentrations, and thecell viability was evaluated with the CellTiter 96 AQueous One SolutionCell Proliferation Assay from Promega Corp. (Madison, Wis., USA). Theindex of cell growth repression ratio was obtained by comparing theOD490 data of treatment group to the negative control. The drug responserate IC₅₀ was calculated with the SPSS 16.0 system. The results areshown in FIG. 7.

For HL-7002 cells, no toxicity was detected for MD-39703 and MD-39433.IC₅₀s of oxaliplatin and 5-FU were 2.44 μM and 4.418 μM, respectively(FIG. 7).

HEK293 kidney cell line cells were treated with drugs (MD-39703,MD-39433, oxaliplatin, or 5-FU) at step-wise concentrations, and thecell viability was evaluated with the CellTiter 96 AQueous One SolutionCell Proliferation Assay from Promega Corp. (Madison, Wis., USA). Theindex of cell growth repression ratio was obtained by comparing theOD490 data of treatment group to the negative control. The drug responserate IC₅₀ was calculated with the SPSS 16.0 system. The results areshown in FIG. 8.

For HEK293, no toxicity was detected for MD-39703 and MD-39433. IC₅₀s ofoxaliplatin and 5-FU were 10.131 μM and 3.744 μM respectively (FIG. 8).

Methods and Strategies:

Cell culture: Colorectal cancer cell lines HCT-116 and HT29 werepurchased from ATCC (Manassas, Va.). The fetal hepatocytes HL-7002 andhuman embryonic kidney cells HEK393 were purchased from ATCC. The cellswere cultured in RPMI+5% Fetal Bovine Serum (FBS).

Drug treatment and cell viability (MTS) assay: The cells (105/100mL/well) were cultured in a 96 well plate, and treated with drugs (e.g.MD-39703, MD-39433, oxaliplatin, or 5-FU) at step-wise concentrationsfrom 0.01 to 300 μM. The cells treated with the solvents were used asthe negative control, and cisplatin and docetaxel were used as thepositive controls. The cells were monitored daily, and the cellviability was evaluated with the Promega CellTiter 96 AQueous OneSolution Cell Proliferation Assay (Promega, Madison, Wis., USA)according to the manufacture manuals. The cell viability was monitoredat OD490 reading in a bio-spectrometer (Perkin Elmer, Walthan, Mass.,USA).

Data analysis: The OD490 reading data were collected hourly from 1 h to4 h after the addition of lysis buffer. The index of cell growthrepression ratio was obtained by comparing the OD490 data of treatmentto the negative control. The drug response rate IC₅₀ was calculated withthe SPSS 16.0.

Summary of Effects:

For colorectal cancer cell line HCT-116, MD-39703 and MD-39433 showedweaker but comparable toxicity than for oxaliplatin and 5-FU. Forcolorectal cancer cell line HT29, MD-39703 and MD-39433 showed a weakerbut comparable toxicity than for oxaliplatin, and stronger toxcity thanfor 5-FU. For HL-7002, an immortalized human fetal hepatic cell line,MD-39703 and MD-39433 showed no toxicity. For HEK293, an immortalizedhuman fetal kidney cell line, MD-39703 and MD-39433 showed no toxicity.

Process to Produce the Co-Crystals and their Charaterizations

Each of the co-crystals of the current invention was formed from aplatinum analogues and a diacid as co-crystal formers. The co-crystalswere obtained by slurrying or grinding platinum analogues and diacids inno solvent or some solvent or mixture of solvents, or by treating thesolution with one or more of several methods, e.g., stepwise heating andcooling the solution, evaporation of the co-crystal formers solution,freeze drying of the co-crystal formers solution, cooling andevaporation of the co-crystal formers solution.

Some crystalline polymorphic forms of the co-crystals of the presentinvention were first produced. Amorphous forms of the co-crystal andother forms may be existent using different methods such as but notlimited to crystallization processes. Polymorphic forms of theco-crystal of platinum analogues were confirmed by X-Ray powderdiffraction (XRPD), thermal gravimetric analysis (TGA) and differentialscanning calorimetry (DSC), scanning electron microscopy (SEM), andother methods. Analysis of the co-crystals showed that each co-crystalcontains one platinum analogue and the corresponding diacid in a 1:1 molratio. Different ratios of the platinum analogues and the acids mayexist using different processes.

Some representative co-crystals with platinum analogues and diacids asco-crystal formers are listed in Table 7.

TABLE 7 Co-crystal Formula number Characterizations

MD36042 HPLC, MS, ¹H-NMR XRPD (FIG. 9); SEM (FIG. 10);

MD39551 HPLC, MS, ¹H-NMR XRPD (FIG. 11); DSC (FIG. 12); SEM (FIG. 13)

MD39433 HPLC, MS, ¹H-NMR XRPD (FIG. 15); SEM (FIG. 16)

MD39442 HPLC, MS, ¹H-NMR XRPD (FIG. 14)

MD39703 HPLC, MS, ¹H-NMR XRPD (FIG. 17); DSC (FIG. 18); SEM (FIG. 19)

HP309 HPLC, MS, ¹H-NMR XRPD (FIG. 20)

MD3176 HPLC, MS, ¹H-NMR XRPD (FIG. 21)

Experiment 1

Mixtures of 550 mg of carboplatin (Pt-01), 300 mg ofcis-endo-5-Norbornene-2,3-dicarboxylic acid (CF-08) and 3.0 mL ofdistilled water were stirred around 30° C. for 5 hours. Then moredistilled water was added to dissolve the mixtures. The obtainedsolution was filtered through a 0.45 um filter and the solution wasdried by stepwise cooling. After cooling dry, the resulted crude crystalwas treated with ethanol and heptane and 448 mg of pure crystal(MD-39551) was obtained. Its characterization by XRPD, DSC/TGA and¹H-NMR confirmed the structure the same as indicated in the Table 6.

Experiment 2

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 400 mg of1,1-cyclobutane dicarboxylate (CF-01 in Table 6) and 3.0 mL of distilledwater were stirred around 30° C. for 5 hours. Then the reaction wascooled to 0-5° C. and stirred over 5 hours. The resulted crude crystalwas obtained by filtering and was washed by cooling distilled water,ethanol and heptane. After dried in vacuum, and 417 mg of pure crystalwas obtained. It was analyzed by HPLC, MS and ¹H-NMR. Thecharacterization indicated 1:1 ratio of lobaplatin to 1,1-cyclobutanedicarboxylate in this co-crystal (MD-3176) structure. Itscharacterization by XRPD, DSC/TGA and ¹H-NMR confirmed the structure thesame as indicated in the Table 6.

Experiment 3

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 520 mg of CF-10A(CF-10A in Table 6) and 3.0 mL of distilled water were stirred around30° C. for 5 hours. Then the reaction was cooled to 0-5° C. and stirredover 5 hours. The resulted crude crystal was obtained by filtering andwas washed by cooling distilled water, ethanol and heptane. After driedin vacuum, and 406 mg of pure crystal was obtained. It was analyzed byHPLC, MS and ¹H-NMR. The characterization indicated 1:1 ratio oflobaplatin to 1,1-cyclobutane dicarboxylate in this co-crystalstructure.

Experiment 4

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 532 mg of CF-10B(CF-10B in Table 6) and 3.0 mL of distilled water were stirred around30° C. for 5 hours. Then the reaction was cooled to 0-5° C. and stirredover 5 hours. The resulted crude crystal was obtained by filtering andwas washed by cooling distilled water, ethanol and heptane. After driedin vacuum, and 375 mg of pure crystal was obtained. It was analyzed byHPLC, MS and ¹H-NMR. The characterization indicated 1:1 ratio oflobaplatin to 1,1-cyclobutane dicarboxylate in this co-crystalstructure.

Experiment 5

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 540 mg of CF-10C(CF-10C in Table 6) and 3.0 mL of distilled water were stirred around30° C. for 5 hours. Then the reaction was cooled to 0-5° C. and stirredover 5 hours. The resulted crude crystal was obtained by filtering andwas washed by cooling distilled water, ethanol and heptane. After driedin vacuum, and 427 mg of pure crystal was obtained. It was analyzed byHPLC, MS and ¹H-NMR. The characterization indicated 1:1 ratio oflobaplatin to 1,1-cyclobutane dicarboxylate in this co-crystalstructure.

Experiment 6

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 540 mg of CF-10D(CF-10D in Table 6) and 3.0 mL of distilled water were stirred around30° C. for 5 hours. Then the reaction was cooled to 0-5° C. and stirredover 5 hours. The resulted crude crystal was obtained by filtering andwas washed by cooling distilled water, ethanol and heptane. After driedin vacuum, and 465 mg of pure crystal was obtained. It was analyzed byHPLC, MS and ¹H-NMR. The characterization indicated 1:1 ratio oflobaplatin to 1,1-cyclobutane dicarboxylate in this co-crystalstructure.

Experiment 7

Mixtures of 400 mg of lobaplatin (Pt-05 in Table 1), 550 mg of Lacticacid and 20.0 mL of distilled water were stirred around 30° C. for 5hours. Then the obtained solution was filtered through 0.45 um filterand the solution was dried by stepwise cooling. After cooling dry, theresulted crude crystal was treated with ethanol and heptane and 355 mgof pure crystal was obtained. It was analyzed by HPLC, MS and ¹H-NMR.The characterization indicated 1:1 ratio of lobaplatin to Lactic acid inthis co-crystal structure.

Experiment 8

General procedure for the preparation of co-crystals based on1,1-cyclobutane dicarboxylate (CF-01 in Table 6) as a co-crystal formerwith one platinum analogue from Pt-02, Pt-03, Pt-04, Pt-05, Pt-06,Pt-07, Pt-08, Pt-09, Pt-10, Pt-11, Pt-12, Pt-13, Pt-14, Pt-15, Pt-16,Pt-17, Pt-18, Pt-19, Pt-20, Pt-21, Pt-22, Pt-23, Pt-24, Pt-25, Pt-26,Pt-27, Pt-28, Pt-29, Pt-30, Pt-31, Pt-32, and Pt-33,

Mixtures of 1.0 mmol of any one platin analogues, 400 mg of1,1-cyclobutane dicarboxylate (CF-01 in Table 6) and 3.0 mL of distilledwater are stirred around 30° C. for 5 hours. Then the reaction is cooledto 0-5° C. and stirred over 5 hours. The resulted crude crystal isobtained by filtering and washed by cooling distilled water, ethanol andheptane. After dried in vacuum, and the desired pure crystal isobtained. Some products are analyzed by XRPD, DSC, HPLC, MS and ¹H-NMR.The characterization indicate 1:1 ratio of selected platin analogues to1,1-cyclobutane dicarboxylate in this co-crystal structure.

Experiment 9

Mixtures of 859 mg of Pt-36 (Pt-31 in Table 5), 1.36 g of1,1-cyclobutane dicarboxylate (CF-01 in Table 6) and 10.0 mL ofdistilled water were stirred around 20° C. for 10 hours. Then thereaction was cooled to 0-5° C. and stirred over 5 hours. The resultedcrude crystal was obtained by filtering and was washed by coolingdistilled water, ethanol and heptane. After dried in vacuum, and 1.15 gof pure crystal was obtained. It was analyzed by HPLC, MS and ¹H-NMR.The characterization indicated 1:1 ratio of Pt-31 (Pt-31 in Table 5) to1,1-cyclobutane dicarboxylate in this co-crystal structure.

Experiment 10

Mixtures of 400 mg of nedaplatin (Pt-03 in Table 1), 520 mg ofphenylmalonic acid (CF-10A in Table 6) and 3.0 mL of distilled waterwere stirred around 30° C. for 5 hours. Then the reaction was cooled to0-5° C. and stirred over 5 hours. The resulted crude crystal wasobtained by filtering and was washed by cooling distilled water, ethanoland heptane. After dried in vacuum, and 470 mg of pure crystal wasobtained. It was analyzed by HPLC, MS and ¹H-NMR. The characterizationindicated 1:1 ratio of nedaplatin to phenylmalonic acid in thisco-crystal structure.

Experiment 11

Mixtures of 410 mg of Pt-06 (Pt-06 in Table 1), 600 mg of1,1-cyclobutane dicarboxylate (CF-01 in Table 6) and 5.0 mL of toluenewas stirred around 30° C. for 5 hours. Then the reaction was cooled to0-5° C. and stirred over 5 hours. The resulted crude crystal wasobtained by filtering and was washed by pre-cooled toluene, ethanol andheptane. After dried in vacuum, and 220 mg of pure crystal was obtained.It was analyzed by HPLC, MS and ¹H-NMR. The characterization indicated1:1 ratio of Pt-06 to 1,1-cyclobutane dicarboxylate in this co-crystalstructure.

Experiment 12

Mixtures of 466 mg of Pt-28 (Pt-21 in Table 5), 520 mg of tartaric acidand 5.0 mL of distilled water was stirred around 30° C. for 5 hours.Then the reaction was cooled to 0-5° C. and stirred over 5 hours. Theresulted crude crystal was obtained by filtering and was washed bypre-cooled distilled water, ethanol and heptane. After dried in vacuum,and 420 mg of pure crystal was obtained. It was analyzed by HPLC, MS and¹H-NMR. The characterization indicated 1:1 ratio of Pt-21 to tartaricacid in this co-crystal structure.

Experiment 13

Mixtures of 580 mg of Pt-32 (Pt-32 in Table 5), 720 mg of phenylmalonicacid (CF-10A in Table 5) and 7.0 mL of distilled water was stirredaround 30° C. for 5 hours. Then the reaction was cooled to 0-5° C. andstirred over 5 hours. The resulted crude crystal was obtained byfiltering and was washed by pre-cooled distilled water, ethanol andheptane. After dried in vacuum, and 437 mg of pure crystal was obtained.It was analyzed by HPLC, MS and ¹H-NMR. The characterization indicated1:1 ratio of Pt-32 to phenylmalonic acid in this co-crystal structure.

Experiment 14

Mixtures of 526 mg of Pt-33 (Pt-28 in Table 5), 520 mg of critic acidand 5.0 mL of distilled water was stirred around 30° C. for 5 hours.Then the reaction was cooled to 0-5° C. and stirred over 10 hours. Theresulted crude crystal was obtained by filtering and was washed bypre-cooled distilled water, ethanol and heptane. The crude solid waspurified by re-crystallization in water. After dried in vacuum, and 320mg of pure crystal was obtained. It was analyzed by HPLC, MS and ¹H-NMR.The characterization indicated 1:1 ratio of Pt-28 to critic acid in thisco-crystal structure.

Experiment 15

Mixtures of 465 mg of Pt-21 (Pt-21 in Table 5), 522 mg of1,2-cis-cyclobutane dicarboxylate (CF-04 in Table 6) and 5.0 mL ofdistilled water was stirred around 20° C. for 7 hours. Then the reactionwas cooled to 0-5° C. and stirred over 10 hours. The resulted crudecrystal was obtained by filtering and was washed by pre-cooled distilledwater, ethanol and heptane. The crude solid was purified byre-crystallization in water. After dried in vacuum, and 458 mg of purecrystal was obtained. It was analyzed by HPLC, MS and ¹H-NMR. Thecharacterization indicated 1:1 ratio of Pt-21 to 1,2-cis-cyclobutanedicarboxylate in this co-crystal structure.

Analytical Methods

X-Ray Powder Diffraction (XRPD):

Polarized light microscopic picture was captured at room temperature(RT). X-ray intensity data were collected at 296(2) K using a BrukerAPEX 11 CCD diffractometer (Mo Kα radiation, λ=0.71073 Å). XRPD patternwas collected by Panalytical Empyrean system at RT. Direct methodsstructure solution, difference Fourier calculations and full-matrixleast-squares refinement against F2 were performed with SHELXTL andOLEX2, See Sheldrick G M. Acta Crystallogr A, 64: 112-122, 2008; and O.V. Dolomanov, et al. J. Appl. Cryst. 42, 339-341, 2009; and Brandenburg,K. DIAMOND, 1999, Crystal Impact GbR, Bonn, Germany. Molecular graphicswere created according to Brandenburg, K. DIAMOND, 1999, Crystal ImpactGbR, Bonn, Germany.

Analytical Instrument: Panalytical Empyrean. The X-ray powderdiffraction was conducted by mounting a sample of the crystallinematerial on a Si single crystal low-background holder and spreading outthe sample into a thin layer with the aid of a microscope slide. The 2θposition was calibrated against Panalytical 640 Si powder standard. Thesample was irradiated with X-rays generated by a copper long-fine focustube operated at 45 kV and 40 mA with a wavelength of Kα1=1.540589angstroms and Kα2=1.544426 angstroms (Kα2/Kα1 intensity ratio is 0.50).The collimated X-ray source was passed through a programmed divergenceslit set at 10 mm and the reflected radiation directed through a 5.5 mmanti-scatter slit. The sample was exposed for 16.3 seconds per 0.013°2-theta increment (continuous scan mode) over the range 3 degrees to 40degrees 2-theta in theta-theta mode. The running time was 3 minutes and57 seconds. The instrument was equipped with a RTMS detector(X'Celerator). Control and data capture was by means of a Dell Optiplex780 XP operating with data collector software.

Persons skilled in the art of X-ray powder diffraction will realize thatthe relative intensity of peaks can be affected by, for example, grainsabove 30 microns in size and non-unitary aspect ratios that may affectanalysis of samples. The skilled person will also realize that theposition of reflections can be affected by the precise height at whichthe sample sits in the diffractometer and the zero calibration of thediffractometer. The surface planarity of the sample may also have alimited effect. Hence the diffraction pattern data presented are notintended to be limited to the absolute values.

Differential Scanning Calorimetry (DSC)

DSC was used as a thermoanalytical method to measure the difference inthe amount of heat required to increase the temperature of a sample andreference was measured as a function of temperature. The general processof DSC is known and the specific instruments and conditions in thefollowing Examples were as follows:

Analytical Instrument: TA Instruments Q2000 DSC;

Heating rate: 10° C. per minute; and Purge gas: nitrogen.

Thermal Gravimetric Analysis (TGA)

TGA was used to measure changes in physical and chemical properties ofsamples as a function of increasing temperature (with constant heatingrate), or as a function of time (with constant temperature and/orconstant mass loss). The general process of TGA is known and thespecific instruments and conditions in the following Examples were asfollows:

Analytical Instrument: TA Instruments Q5000 TGA;

Heating rate: 10° C. per minute; and

Purge gas: nitrogen.

Sample Pharmaceutical Composition and its Administration

Aqueous or solid pharmaceutical composition of the present inventioncomprises an effective amount of the co-crystal of the currentinvention, e.g. MD39551, with or without an appropriate amount of atleast one additional therapeutic agent or adjuvant therapy agent. Theco-crystal, as well as the therapeutic agent or adjuvant therapy agent,may be dissolved or dispersed in a pharmaceutical acceptable carrier oraqueous media.

Depending on the particular cancer to be treated, administration ofpharmaceutical composition according to the present invention can viaany common route as long as the target issue is available via the route.For example, the pharmaceutical composition may be administered byinfusion, injection, or via the oral route.

A Number of Pharmaceutical Compositions were Produced:

Pharmaceutical composition sample A: 70 g of MD-39551 was dissolved inpre-treated normal saline or 5% of aqueous glucose (in water) and thefinal volume of the solution was adjusted to 5.0 L. Then the solutionwas filtered through 0.22 um filter and dispersed into ample bottleswith 50.0 mL in each.

1-31. (canceled)
 32. A co-crystal comprising a platinum analogue offormula Pt-02:

and a diacid selected from the group consisting of:


33. The co-crystal of claim 32, wherein the diacid is selected from thegroup consisting of formulas CF-01, CF-02, and CF-08.
 34. The co-crystalof claim 32, which is a co-crystal comprising the platinum analogue offormula Pt-02 and the diacid of formula CF-02 and having an XRPD patterncomprising peaks at diffraction angles 2-Theta of 7.3°, 9.4°, 10.1°,12.5°, 13.6° and 23.4°±0.2, an XRPD pattern comprising peaks as setforth in FIG. 14 or an XRPD pattern substantially similar to the patternas set forth in FIG. 14; or a co-crystal comprising the platinumanalogue of formula Pt-02 and the diacid of formula CF-08 and having anXRPD pattern comprising peaks at diffraction angles 2-Theta of 7.9°,11.9°, 14.5°, 15.8°, 17.0°, 17.4° and 17.8°±0.2, an XRPD patterncomprising peaks as set forth in FIG. 17 or an XRPD patternsubstantially similar to the pattern as set forth in FIG.
 17. 35. Theco-crystal of claim 32, which is a co-crystal comprising the platinumanalogue of formula Pt-02 and the diacid of formula CF-01 and having anXRPD pattern comprising peaks at diffraction angles 2-Theta of 7.1°,9.2°, and 10.1°±0.2.
 36. The co-crystal of claim 32, which is aco-crystal comprising the platinum analogue of formula Pt-02 and thediacid of formula CF-08 and having an XRPD pattern comprising peaks asset forth in FIG. 15 or an XRPD pattern substantially similar to thepattern as set forth in FIG.
 15. 37. A pharmaceutical compositioncomprising the co-crystal of claim
 32. 38. A pharmaceutical compositioncomprising the co-crystal of claim
 34. 39. A pharmaceutical compositioncomprising the co-crystal of claim
 35. 40. A pharmaceutical compositioncomprising the co-crystal of claim
 36. 41. The pharmaceuticalcomposition of claim 37, which is an aqueous composition, wherein theco-crystal is dissolved or dispersed in a pharmaceutically acceptablecarrier or aqueous media.
 42. The pharmaceutical composition of claim37, further comprising a therapeutic agent or adjuvant therapy agentselected from the group consisting of folic acid, coenzyme Q10,curcumin, glutathione (GSH), aloe vera, oryzanol, 5-fluorouracil, andbortezomib.
 43. A method of treating cancer in a subject in needthereof, comprising administering to the subject the pharmaceuticalcomposition of claim 37, wherein the co-crystal is in a therapeuticallyeffective amount.
 44. A method of treating cancer in a subject in needthereof, comprising administering to the subject the pharmaceuticalcomposition of claim 39, wherein the co-crystal is in a therapeuticallyeffective amount.
 45. A method of treating cancer in a subject in needthereof, comprising administering to the subject the pharmaceuticalcomposition of claim 40, wherein the co-crystal is in a therapeuticallyeffective amount.
 46. The method of claim 43, wherein the cancer isprostate cancer, colorectal cancer, or renal adenocarcinoma.
 47. Amethod of preparing a co-crystal comprising: a) mixing a platinumanalogue and a diacid in water, b) slurrying or stirring the mixturefrom step a) for a sufficient period of time to form a co-crystal of theplatinum analog and the diacid; and optionally c) isolating theco-crystal, wherein the platinum analog is a platinum analogue offormula Pt-02:

and the diacid is selected from the group consisting of:


48. The method of claim 47, wherein the molar ratio of the platinumanalogue to the diacid is in range of 1:0.1 to 1:20.
 49. The method ofclaim 48, wherein the diacid is CF-01.
 50. The co-crystal produced bythe method of claim
 47. 51. The co-crystal produced by the method ofclaim 49.